Validation of taxon names described in Deltocephalinae from China (Hemiptera: Cicadellidae).

The purpose of this paper is to validate 4 generic names and 34 specific names previously introduced in the Deltocephalinae from China (Hemiptera: Cicadellidae) by Li, Dai & Xing (2011). The names are unavailable in the original publication, because of the missing information on the type specimen repository. The following genera and species are introduced as new in this publication: Bambusananus, Li & Xing; Branchana Li; Cyclevertex Li & Xing; Lineana Li & Xing; Abrus biprocessus Li; Abrus graciaedeagus Li; Bambusana biflaka Li; Bambusana nigrimaculata Li; Bambusananus furcatus Li & Xing; Branchana xanthota Li; Cyclevertex furcatum Li & Xing; Lineana albipunctata Li & Xing; Lineana ductaedeagusa Li & Xing; Protensus nigrifrons Li & Xing; Protensus yanheensis Li & Xing; Paramesodes menghaiensis Xing & Li; Yuanamia producta (Xing & Li, 2011); Hishimonoides brevis Li, 2011; Litura tripunctata Li; Changwhania huajiangensis Xing & Li; Elginus productus Xing & Li; Metalimnus maoershanensis Xing & Li; Nicolaus bihamatus Xing & Li; Paralaevicephalus bisubulatus Xing & Li; Paralaevicephalus brevissimus Xing & Li; Paralaevicephalus spinosus Xing & Li; Scaphoideus annulatus Li; Scaphoideus bannaensis Li; Longicornus longus Xing & Li; Melanetettix mengyangensis Xing & Li; Osbornellus aurantius Xing & Li; Osbornellus suiyangensis Xing & Li; Osbornellus conoideus Xing & Li; Phlogothamnus acutaedeagus Li; Phlogothamnus luteoguttatus Li; Phlogothamnus productus Li & Xing; Phlogothamnus rugosus Li; Phlogotettix fanjingshanensis Li.

Optimal cold storage protocol for Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae), a biological control agent for coleopteran pests in stored products.

BACKGROUND: Anisopteromalus calandrae (Howard) is a solitary ectoparasitoid with wide-ranging potential applications as a natural biological control agent against various coleopterous pests in food warehouses. Implementing an effective cold storage program is crucial for extending the shelf life of biological control agents and ensuring their stable and abundant supply. Herein, we attempted to determine the optimal cold storage conditions for Anisopteromalus calandrae by investigating the effect of cold storage at three different temperatures (7, 13, and 19 °C) for 7, 21, and 35 days on four developmental stages (late-instar larvae, early-stage pupae, mid-stage pupae, and 2-day-old adults). Additionally, we explored the maximum cold storage potential by observing early-stage pupae stored at 13 °C for various durations (30, 60, 90, 120, and 150 days). RESULTS: The most suitable cold storage temperature for the early-stage pupae of Anisopteromalus calandrae was 13 °C, and the highest adult emergence rate (98.3%) was after 90 days of storage at 13 °C. Furthermore, we did not find any significant effect on longevity (female: 44.3 days; male: 38.1 days) or fecundity (121.7 wasps). The female ratio ranged from 43.5% to 50.8%. More importantly, cold storage did not adversely affect the developmental duration or fecundity of the offspring. CONCLUSION: This study offers crucial insights for managing Anisopteromalus calandrae populations under laboratory conditions and lays the foundation for potential industrial production and development. © 2023 Society of Chemical Industry.

Identification and immune analysis of antimicrobial peptides from the cigarette beetle (Lasioderma serricorne).

Antimicrobial peptides (AMPs) in insects are endogenous peptides that are effector components of the innate defense system of the insect. AMPs may serve as antimicrobial agents because of their small molecular weight and broad-spectrum antimicrobial activity. In this study, we performed transcriptome analysis of cigarette beetle (Lasioderma serricorne) larvae, parasitized by the ectoparasitic wasp, Anisopteromalus calandrae. Several AMP genes were significantly upregulated following A. calandrae parasitism, postulating the hypothesis that the parasitization enhanced the host's resistance against pathogenic microorganisms through the regulation of host AMP genes. Specifically, 3 AMP genes (LsDef1, LsDef2, and LsCole) were significantly upregulated and we studied their immune function in L. serricorne. Immune challenge and functional analysis showed that LsCole was responsible for the immune response against Gram-negative and Gram-positive bacteria, while LsDef1 and LsDef2 were involved in insect defense against Gram-positive bacteria. Purified recombinant LsCole exhibited antimicrobial activities against the Gram-negative bacterium Escherichia coli and the Gram-positive bacterium Staphylococcus aureus. LsDef2 showed an antibacterial effect against S. aureus. LsCole and LsDef2 exhibited antibiofilm activity against S. aureus. The 2 AMPs disrupted cell membranes and caused leakage of S. aureus cell contents. The results indicated that the 3 AMPs in L. serricorne are involved in the innate immunity of this pest insect. These AMPs may have potential as antimicrobial agents for bacterial infection chemotherapy. Hence, data are discussed in relation to new control strategies with greater biosafety against pest insects with use of microbial biocontrol agents in combination with RNA interference against the insect's defensive AMP genes.

Comparative analysis of codon usage patterns and phylogenetic implications of five mitochondrial genomes of the genus Japanagallia Ishihara, 1955 (Hemiptera, Cicadellidae, Megophthalminae).

Japanagallia is a genus of Cicadomorpha in the family of leafhoppers that are plant piercing-sucking insects, and it is difficult to distinguish by morphological characteristics. So far, only one complete mitochondrial genome data has been reported for the genus Japanagallia. Therefore, in order to better understand this group, we assembled and annotated the complete mitochondrial genomes of five Japanagallia species, and analyzed their codon usage patterns. Nucleotide composition analysis showed that AT content was higher than GC content, and the protein-coding sequences preferred to end with A/T at the third codon position. Relative synonymous codon usage analysis revealed most over-represented codon ends with A or T. Parity plot analysis revealed the codon usage bias of mitochondrial genes was influenced by both natural selection and mutation pressure. In the neutrality plot, the slopes of regression lines were < 0.5, suggesting that natural selection was playing a major role while mutation pressure was of minor importance. The effective number of codons showed that the codon usage bias between genes and genomes was low. Correspondence analysis revealed that the codon usage pattern differed among 13 protein-coding genes. Phylogenetic analyses based on three datasets using two methods (maximum likelihood and Bayesian inference), restored the Megophthalminae monophyly with high support values (bootstrap support values (BS) = 100, Bayesian posterior probability (PP) = 1). In the obtained topology, the seven Japanagallia species were clustered into a monophyletic group and formed a sister group with Durgade. In conclusion, our study can provide a reference for the future research on organism evolution, identification and phylogeny relationships of Japanagallia species.

Six new species of the leafhopper subgenus Pediopsoides (Pediopsoides) (Hemiptera, Cicadellidae, Eurymelinae, Macropsini) from China.

The nominate subgenus Pediopsoides (Pediopsoides) Matsumura, 1912 is widely distributed in the Oriental region but has high species diversity only in southern China. The present paper describes and illustrates six new species of Pediopsoides (Pediopsoides) namely, P. (P.) ailaoshanensis Li & Dai, sp. nov., P. (P.) quadrispinosus Li & Dai, sp. nov., P. (P.) flavus Li & Dai, sp. nov., and P. (P.) pianmaensis Li & Dai, sp. nov. all collected in Yunnan Province, southwestern China, P. (P.) maoershanensis Li & Dai, sp. nov., found in Guangxi Autonomous Region, southern China, and P. (P.) huangi Li & Dai sp. nov., from Taiwan, previously incorrectly recorded as a new name by Li & Dai, 2018 in Dai et al. 2018: 203 for P. (P.) femorata Huang & Viraktamath, 1993 (nec Pediopsisfemorata Hamilton, 1980). Two new junior synonyms of Sispocnis Anufriev, 1967 are proposed, i.e., Digitalis Liu & Zhang, 2002, syn. nov. and Neosispocnis Dmitriev, 2020, syn. nov.

Structural Characteristics and Phylogenetic Analysis of the Mitochondrial Genomes of Four Krisna Species (Hemiptera: Cicadellidae: Iassinae).

Krisna species are insects that have piercing-sucking mouthparts and belong to the Krisnini tribe in the Iassinae subfamily of leafhoppers in the Cicadellidae family. In this study, we sequenced and compared the mitochondrial genomes (mitogenomes) of four Krisna species. The results showed that all four mitogenomes were composed of cyclic double-stranded molecules and contained 13 protein-coding genes (PCGs) and 22 and 2 genes coding for tRNAs and rRNAs, respectively. Those mitogenomes exhibited similar base composition, gene size, and codon usage patterns for the protein-coding genes. The analysis of the nonsynonymous substitution rate (Ka)/synonymous substitution rate (Ks) showed that evolution occurred the fastest in ND4 and the slowest in COI. 13 PCGs that underwent purification selection were suitable for studying phylogenetic relationships within Krisna. ND2, ND6, and ATP6 had highly variable nucleotide diversity, whereas COI and ND1 exhibited the lowest diversity. Genes or gene regions with high nucleotide diversity can provide potential marker candidates for population genetics and species delimitation in Krisna. Analyses of parity and neutral plots showed that both natural selection and mutation pressure affected the codon usage bias. In the phylogenetic analysis, all subfamilies were restored to a monophyletic group; the Krisnini tribe is monophyletic, and the Krisna genus is paraphyletic. Our study provides novel insights into the significance of the background nucleotide composition and codon usage patterns in the CDSs of the 13 mitochondrial PCGs of the Krisna genome, which could enable the identification of a different gene organization and may be used for accurate phylogenetic analysis of Krisna species.

Interaction Analysis of lncRNA and mRNA Based on the Full-Length Transcriptome of the Nymph-to-Adult Developmental Transition of Sogatella furcifera.

Little is known on how long noncoding RNAs (lncRNAs) and mRNAs cooperatively participate in regulating the nymph-to-adult development transition of Sogatella furcifera. Herein, lncRNA and mRNA libraries were constructed in three different developmental stages of S. furcifera, namely, prior to (PE), during (DE), and after (AE) ecdysis. Overall, 4649 lncRNAs were identified and divided into intergenic (53.90%), intronic (1.33%), sense (8.99%), antisense (21.75%), and bidirectional (3.94%) lncRNAs. Moreover, 795 differentially expressed lncRNAs were identified. Specifically, upon comparing PE and DE, 2719 target mRNAs were predicted for 574 lncRNAs. Upon comparing PE and AE, 2816 target mRNAs were predicted for 627 lncRNAs. Finally, upon comparing DE and AE, 51 target mRNAs were predicted for 35 lncRNAs. Kyoto Encyclopedia of Genes and Genome functional enrichment analysis revealed that the target genes of 795 lncRNAs were enriched in metabolic pathways, amino sugar and nucleotide sugar metabolism, and fatty acid metabolism. Subsequently, interaction analysis indicated that MSTRG.16086.1, MSTRG.16087.1, and MSTRG.2447.1 were functionally associated with cuticle protein and chitin biosynthesis. Finally, 11 differentially expressed lncRNAs were significantly enriched in 3rd and 4th instar nymphs. Our findings suggest that lncRNAs play a critical regulatory role during the molting of S. furcifera.

Description of the whole mitochondrial genome of Bhatia longiradiata (Hemiptera, Cicadellidae, Deltocephalinae: Drabescini) and phylogenetic relationship.

BACKGROUND: Mitochondrial genomes are extremely conserved in genetic processes and valuable molecular indications for phylogenetic and evolutionary examination, but the mitochondrial genome of Bhatia has not yet been reported. OBJECTIVE: The target of this writing was to clarify the structural module of the mitochondrial genes of Bhatia longiradiata, verify the monophyletic of Drabescini, and explore the phylogenetic relationship between Drabescini with other leafhoppers. METHODS: We performed sequencing and explanatory note of the mitochondrion of Bhatia longiradiata. The phylogeny relation was created by ML and Bayesian approaches using three dissimilar datasets (PCG12, PCG12rRNA, and AA), which were constructed to discuss the phylogenetic status of Bhatia longiradiata. RESULTS: To report the architectural feature of the chondriosome of Bhatia longiradiata is a seal double-stranded annular molecule with 16,122 bp measurement and cover typically 37 genes. Several tandem repetitive units were observed in an AT enrichment area. The analysis showed that the branching relationships among the six trees were generally consistent, and each of the subfamilies was individually clustered into a monophyletic group within Cicadellidae. Bhatia longiradiata and other members of the Drabescini were aggregated into a clade that was situated within the Deltocephalinae. CONCLUSION: The mitochondrial genome of Bhatia longiradiata covers 37 typical genes and a control region, which covers six tandem repeats. All species of Drabescini procedure a clade within Deltocephalinae. Drabescini and Scaphoideini form a branch and show a sister relationship with strong support. Therefore, we support the relegation of Selenocephalinae to a clan within Deltocephalinae.

Immune function analysis of LsSd, a transcription factor of the Hippo signaling pathway, in the cigarette beetle Lasioderma serricorne.

The Scalloped (Sd) is a transcription factor that regulates organ size control in the Hippo-signaling pathway. Recent studies have showed that Hippo signaling also functions in the innate immune response. Although the Sd gene has been reported in many insects, their immune functions remain unexplored. In this study, the LsSd gene of Lasioderma serricorne, with a complete open reading frame that encodes a protein composed of 402 amino acids was identified. LsSd was predominantly expressed in early pupae. Tissue-specific analyses revealed that the highest concentrations of LsSd were detected in the midgut and brain. At 1-24 h after Escherichia coli infection, LsSd expression increased substantially. However, LsSd expression was downregulated 3-12 h after Staphylococcus aureus infection. RNA interference-mediated silencing of the LsSd transcript resulted in deformed, considerably smaller, and degenerated wings. Meanwhile, LsCycE, LsDiap1, and LsVg, which are involved in cell proliferation and wing development, were drastically reduced when LsSd was depleted. In a survival assay, the LsSd knockdown considerably decreased the susceptibility to S. aureus, a gram-positive bacterium. In addition, knockdown of LsSd remarkably downregulated the transcription of LsCact in response to S. aureus stimulation, while upregulating the expression of five immune-related genes. Our results provide conclusive proof for the important roles of LsSd in the immune response of L. serricorne.

Mitogenomic phylogeny of Typhlocybinae (Hemiptera: Cicadellidae) reveals homoplasy in tribal diagnostic morphological traits.

The subfamily Typhlocybinae is a ubiquitous, highly diverse group of mostly tiny, delicate leafhoppers. The tribal classification has long been controversial and phylogenetic methods have only recently begun to test the phylogenetic status and relationships of tribes. To shed light on the evolution of Typhlocybinae, we performed phylogenetic analyses based on 28 newly sequenced and 19 previously sequenced mitochondrial genomes representing all currently recognized tribes. The results support the monophyly of the subfamily and its sister-group relationship to Mileewinae. The tribe Zyginellini is polyphyletic with some included genera derived independently within Typhlocybini. Ancestral character state reconstruction suggests that some morphological characters traditionally considered important for diagnosing tribes (presence/absence of ocelli, development of hind wing submarginal vein) are homoplastic. Divergence time estimates indicate that the subfamily arose during the Middle Cretaceous and that the extant tribes arose during the Late Cretaceous. Phylogenetic results support establishment of a new genus, Subtilissimia Yan & Yang gen. nov., with two new species, Subtilissimia fulva Yan & Yang sp. nov. and Subtilissimia pellicula Yan & Yang sp. nov.; but indicate that two previously recognized species of Farynala distinguished only by the direction of curvature of the processes of the aedeagus are synonyms, that is, Farynala dextra Yan & Yang, 2017 equals Farynala sinistra Yan & Yang, 2017 syn. nov. A key to tribes of Typhlocybinae is provided.

Descriptions of two new Idiocerini leafhoppers of the genus Idioscopus (Hemiptera, Cicadellidae) from China.

Two new species of the leafhopper genus Idioscopus Baker are described from China: Idioscopusbihamulus sp. nov. and I.ventrispinus sp. nov., the latter recorded on a species of Myrica L. (Myricaceae) as its host plant. A key and checklist to species of the genus from China are provided and Idioscopustaiwanus Huang & Maldonado-Capriles, 1992 is placed as a junior synonym of Idioscopusclypealis (Lethierry, 1889), syn. nov.

Comparison of mitogenomes of three Petalocephala species (Hemiptera: Cicadellidae: Ledrinae) and their phylogenetic analysis.

Ledrinae is a unique group of leafhoppers with a distinct appearance. Petalocephala is the largest Ledrinae genus that is difficult to identify except by dissecting the male genitals. To date, research on Ledrinae is relatively less compared with other leafhoppers. Therefore, to better understand this group, we sequenced and analyzed three complete Petalocephala mitochondrial genomes. We comparatively analyzed these general Petalocephala genomic features (including size, AT content, AT/GC skew, 13 protein-coding gene nucleotide compositions, etc.), and predicted 22 transfer RNA secondary structures. We obtained highly consistent phylogenetic results within Cicadellidae based on mitogenomic data using the maximum likelihood and Bayesian methods. Our results showed that all subfamilies were monophyletic and had a high node support rate, and there was a sister group relationship between Ledrinae and all other leafhopper groups. Furthermore, treehoppers were found to originate from leafhoppers and showed sister group relationships with Megophthalminae. Within Ledrinae, all phylogenetic trees supporting phylogenetic relationships were as follows: ([P. dicondylica + P. gongshanensis] + [Tituria pyramidata + [Ledra auditura + P. gongshanensis]]) Based on the complete mitogenome phylogenetic analysis and the comparison of morphological characteristics, we propose that Petalocephala is not monophyletic.

Description of two new species of the genus Siniassus (Hemiptera: Cicadellidae: Iassinae: Hyalojassini) from China.

Two new species of leafhoppers: Siniassus gracilitus sp. nov. and S. nigrimaculatus sp. nov., of tribe Hyalojassini, subfamily Iassinae (Hemiptera: Cicadellidae) are described and illustrated from China. A key to Siniassus species and updated checklist with distributions are provided.

Description of a new species of the leafhopper genus Chinaocerus (Hemiptera: Cicadellidae) from Yunnan, China.

A new species Chinaocerus dentaedeagus sp. nov. is described and illustrated from Yunnan, China. The new species can be distinguished from other species of the genus Chinaocerus by aedeagal shaft subapex with pair of dentate lamellar. Meanwhile, a checklist of the genus and a key to the genera from China are provided.

Description of three new species of the leafhopper genus Idioscopus Baker, (Hemiptera: Cicadellidae: Eurymelinae) from Yunnan, China.

Three new species of the leafhopper genus Idioscopus Baker, 1915 from China: I. furcaprocessus sp. nov., I. longiprocessus sp. nov., and I. serratastylus sp. nov. are described and photographed based on specimens collected in Yunnan Province of southern China. These three new species can be recognised mainly by the coloration and male genitalia.

Chromosome-Level Genome Assembly of Nephotettix cincticeps (Uhler, 1896) (Hemiptera: Cicadellidae: Deltocephalinae).

The green rice leafhopper, Nephotettix cincticeps (Uhler), is an important rice pest and a vector of the rice dwarf virus in Asia. Here, we produced a high-quality chromosome-level genome assembly of 753.23 Mb using PacBio (∼110×) and Hi-C data (∼94×). It contained 163 scaffolds and 950 contigs, whose scaffold/contig N50 lengths reached 85.36/2.57 Mb. And 731.19 Mb (97.07%) of the assembly was anchored into eight pseudochromosomes. Genome completeness was attained to 97.0% according to the insect reference Benchmarking Universal Single-Copy Orthologs (BUSCO) gene set (n = 1,367). We masked 347.10 Mb (46.08%) of the genome as repetitive elements. Nine hundred sixty-two noncoding RNAs were identified and 14,337 protein-coding genes were predicted. We also assigned GO term and KEGG pathway annotations for 10,049 and 9,251 genes, respectively. Significantly expanded gene families were primarily involved in immunity, cuticle, digestion, detoxification, and embryonic development. This study provided a crucial genomic resource for better understanding on the biology and evolution in family Cicadellidae.

Structural features of the mitogenome of the leafhopper genus Cladolidia (Hemiptera: Cicadellidae: Coelidiinae) and phylogenetic implications in Cicadellidae.

The first two complete mitogenomes of the leafhopper genus Cladolidia (C. biungulata and C. robusta) were sequenced and annotated to further explore the phylogeny of Cladolidia. Both the newly sequenced mitogenomes have a typical circular structure, with lengths of 15,247 and 15,376 bp and A + T contents of 78.2% and 78%, respectively. We identified a highly conserved genome organization in the two Cladolidia spp. through comparative analysis that included the following assessments: genome content, gene order, nucleotide composition, codon usage, amino acid composition, and tRNA secondary structure. Moreover, we detected the base heterogeneity of Cicadellidae mitogenomic data and constructed phylogenetic trees using the nucleotide alignments of 12 subfamilies of 58 leafhopper species. We noted a weak heterogeneity in the base composition among the Cicadellidae mitogenomes. Phylogenetic analyses showed that the monophyly of each subfamily was generally well supported in the family Cicadellidae; the main topology was as follows: (Deltocephalinae + (Treehoppers + ((Megophthalminae + (Macropsinae + (Hylicinae + (Coelidiinae +Iassinae)) + (Idiocerinae + (Cicadellinae + (Typhlocybinae + (Mileewinae + (Evacanthinae +Ledrinae)))))))))). Within Coelidiinae, phylogenetic analyses revealed that C. biungulata and C. robusta belong to Coelidiinae and the monophyly of Cladolidia is well supported. In addition, on the basis of complete mitogenome phylogenetic analysis and the comparison of morphological characteristics, we further confirm the genus Olidiana as a paraphyletic group, suggesting that the genus may need taxonomic revisions.

The complete mitochondrial genome of Oenopia sauzeti (Coleoptera: Coccinellidae).

Oenopia sauzeti Mulsant are predators of various aphids, such as Myzus persicae, Rhopalosiphum maidis, and Sitobion avenae. We sequenced the mitochondrial genome (mitogenome) of O. sauzeti. Consistent with other mitogenomes of Coleoptera, the circular mitogenome of O. sauzeti consists of 17,630 bp, including 13 PCG genes, 2 rRNA genes, and 22 tRNA genes. The mitogenome differs in 2 long non-coding regions of 1285 bp and 1843 bp. The 22 tRNA genes products are folded into the typical cloverleaf secondary structure, except trnS1, which lacks the dihydrouridine (DHU) arm. The base composition is AT-biased (80.1%). The phylogenetic tree confirms monophyly at the genus level within the Coccinellinae and supports O. sauzeti as a sister group to Aiolocaria hexaspilota.

Mitogenomics of five Olidiana leafhoppers (Hemiptera: Cicadellidae: Coelidiinae) and their phylogenetic implications.

Similar morphological characteristics and limited molecular data of Olidiana resulted in their unknown phylogenetic statuses and equivocal relationships. To further understand the genus Olidiana, we sequenced and annotated five Olidiana complete mitochondrial genomes (mitogenomes). Our results show that Olidiana mitogenomes range from 15,205 bp to 15,993 bp in length and include 37 typical genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) and a control region. Their nucleotide composition, codon usage, features of control region, and tRNA secondary structures are similar to other members of Cicadellidae. We constructed the phylogenetic tree of Cicadellidae using the maximum likelihood (ML) and Bayesian inference (BI) methods based on all valid mitogenome sequences. The most topological structure of the obtained phylogenetic tree is consistent. Our results support the monophyletic relationships among 10 subfamilies within Cicadellidae and confirm Iassinae and Coelidiinae to be sister groups with high approval ratings. Interestingly, Olidiana was inferred as a paraphyletic group with strong support via both ML and BI analyses. These complete mitogenomes of five Olidiana species could be useful in further studies for species diagnosis, evolution, and phylogeny research within Cicadellidae.

Two new species of the leafhopper genus Calodia Nielson (Hemiptera, Cicadellidae, Coelidiinae) from China, with a checklist and key to Chinese species.

Two new species of the leafhopper genus Calodia Nielson are described and illustrated: C. quadrimacula sp. nov. from Guizhou and Yunnan Provinces and C. zuoae sp. nov. from Yunnan Province, China. A checklist along with distribution and a key to species based on male genitalia of the genus Calodia from China are provided. Olidiana nigritibiana (Li), comb. nov. (earlier in the genus Calodia) is proposed. At present, this genus comprises 45 known species worldwide, of which 19 species are recorded from China.